2025-08-28
ENA first public
ERC000011
ENA-CHECKLIST
Princess Maxima Center for Pediatric Oncology
INSDC center name
public
INSDC status
ERS24550422
SRA accession
E-MTAB-15147:Sample_timecourse_7
Submitter Id
ArrayExpress
broker name
WA09
cell line
inferred
cell type
not collected
collection date
human
common name
Protocols: Multiple organoids from the same batch and age were pooled. For timepoints after Matrigel embedding, Cell Recovery Solution (Corning, 354253) was used to dissolve the Matrigel. Organoids were incubated in this solution at 4°C for 15 minutes, halved, and washed in HBSS without Ca²⁺ and Mg²⁺. A single cell supsension was obtained using the Neural Tissue Dissociation Kit (Miltenyi Biotec, 130-092-628) according manufactures protocol.  Sequencing samples were thawed by warming the cryovials in a 37°C water bath until only a small clump of ice remained. The contents were then transferred to 10 mL of pre-warmed DMEM containing 10% FBS and centrifuged to pellet the cells. The cells were washed twice with PBS containing 5% BSA and filtered through a 40 µm Flowmi cell strainer to remove debris and aggregates. Approximately 30,000 dissociated cells per sample/timepoint were loaded into separate channels of a single Chromium controller (10x Genomics) chip. Consequently, gel bead-in-emulsions (GEMS) were generated using 10X Genomics Single Cell 3’ Reagent Kit v4. Sequencing libraries were prepared using Single Cell 3’ Reagent Kits v4 (10X Genomics) and then converted using the NovaSeq 6000 S2 Reagent Kit (Illumina). Library generation was provided by Leiden Genome Technology Center (Leiden, The Netherlands)
description
blastula stage
developmental stage
normal
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
blastocyst
organism part
Homo sapiens
scientific_name
female
sex
Sample_timecourse_7
title